A multiparameter fiber optic biosensor for continuous determination of cholesterol and glucose was developed. This sensor was based on poly(N-isopropylacrylamide) (PNIPAAm)-immobilized glucose oxidase (GOx) complex (PIGC) and immobilized cholesterol oxidase (COD). The immobilized COD catalysis to the oxidation of cholesterol and PIGC catalysis to the oxidation of glucose could be performed at different temperatures. Therefore, the sensor could detect cholesterol and glucose continuously by changing temperature. The optimal detection conditions for glucose were achieved with pH 6.5, 30 degrees C, and 10 mg GOx (in 100-mg carrier), and those for cholesterol were achieved with pH 7.5, 33 degrees C, and 25 mg COD (in 250-mg carrier). The sensor has the cholesterol detection range of 20-250 mg/dL and the glucose detection range of 50-700 mg/dL. This biosensor has outstanding repeatability and selectivity, and the detection results of the practical samples are satisfactory.